Streptavidin two-dimensional (2D) crystals were grown on mica-supported phospholipid bilayers containing a biotinylated phospholipid. Their topography structure obtained by atomic force microscopy compares favorably with the electron microscopy analysis of streptavidin 2D crystals grown on lipid monolayers at the air-water interface. The streptavidin 2D crystals were used as a functionalized matrix for anchoring phospholipid bilayers on top of them. Biotinylated liposomes were found to bind specifically to the streptavidin matrix and to transform, locally, into flat lipid surfaces. Height measurements as well as parallel electron microscopy studies performed with biotinylated lipid tubules suggest that the flat lipid surfaces are indeed single lipid bilayers anchored to the ordered streptavidin matrix.